DEV01886 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 (Adobe PDF) -

  Fig. S1. Structure of the genomic region of novel BM protein-trap lines. The position of the GFP exon (shown as a green bar) was determined by sequencing of protein-trap genomic DNA, and is represented on the annotated genomic region (http://flybase.bio.indiana.edu). (A) Protein-trap line G205 is inserted at position 83193 of scaffold AE003608, in the second and biggest intron of the viking gene (collagen IV a 2 chain). The GFP-Vkg protein-trap line used throughout the study (vkg^G454) is inserted 963 bp upstream of G205 (position 84156 of scaffold AE003608; data not shown) (Morin et al., 2001). The size of the resulting GFP fusion protein has been determined by western-blot using anti-GFP antibodies (arrowhead); GFP-G205 migrates at the same position as vkg^G454. Asterisk represents non-specific staining. (B) Protein-trap lines G6, 658, 1700, 1973, 2840 and 2867 are all inserted in between CG12497 and CG7981/Trol/Perlecan gene at the following positions of genomic scaffold AE003424: 158962 (G6), 158957 (1700), 158559 (1973), 158959 (2840), 158957 (2867; same insertion site as 1700). The high molecular weight (>400 kDa) of the GFP fusion proteins, as determined by western-blot (arrowhead), indicates that the protein-trap lines have targeted the perlecan protein, whose N-terminal region is still not determined (current calculated molecular weight of perlecan is 355-466 kDa depending on the isoform; see FlyBase at http://flybase.bio.indiana.edu/.bin/fbidq.html?FBgn0001402&resultlist=fbgn17958.data

• Movie 1 -

  Movie 1. 3D-reconstructed anterior region of the egg chamber shown in Fig. 3A-C. Nuclei are shown in blue (Hoeschst), polar cells are in red (Fas2 staining) and GFP-Vkg is in green. Anterior is up.

• Movie 2 -

  Movie 2. Same egg chamber as shown in Movie 1. Only polar cells (white) and the apical cap (blue) are shown. Anterior is up.